Serial Propagation of the Microsporidian Enterocytozoon bieneusi of Human Origin in Immunocompromised Rodents

Abstract

Enterocytozoon bieneusi , a microsporidian, is clinically one of the most significant opportunistic causes of diarrhea and wasting associated with profound human immunodeficiencies. The lack of an animal model for E. bieneusi hinders serious investigations and limits the availability of spores to individuals with severe human immunodeficiency virus/AIDS disease who are infected with E. bieneusi . The development of procedures for purification and concentration of spores from stools of infected humans has led to the production of immune reagents and provided a source of spores to conduct research, including attempts to develop and serially propagate E. bieneusi in rodent models. We have evaluated and successfully infected six different immunodeficient and/or immunosuppressed rodent models and have demonstrated persistent infections lasting at least 18 weeks in SCID mice and in nude rats. To enhance the intensity and duration of infection in these two models, animals were given anti-gamma interferon monoclonal antibody injections at regular intervals. Of the six models evaluated, nude rats and gerbils immunosuppressed with dexamethasone excreted the highest number of spores and for longer time periods. Four different E. bieneusi isolates were equally infectious, and one of them was serially propagated in nude rats six times over a period of 10 months. Typically, rats challenged orally with 10 ⁴ spores yielded 2 × 10 ⁷ to 6.3 × 10 ⁷ spores per single fecal sample when the level of spores was measured 2 weeks later. Rodent models and a nonhuman source of fresh spores will considerably enhance future investigations on this important opportunistic pathogen, including the screening and evaluation of urgently needed chemotherapeutic agents.